Submitted December 30, 2008
Accepted February 14, 2009
Antisense-mediated exon skipping to correct IL-12R
1 deficiency in T cells
Esther van de Vosse*, Els M. Verhard, Roelof A. de Paus, and Jaap T. van Dissel
Department of Infectious Diseases, Leiden University Medical Center, Leiden, Netherlands
* Corresponding author; email: e.van_de_vosse{at}lumc.nl.
Patients with Mendelian Susceptibility to Mycobacterial Disease (MSMD) suffer from severe, recurrent life-threatening infections with otherwise poorly-pathogenic mycobacteria and salmonellae. The extreme susceptibility is due to genetic defects in the IL-12/IFN-
pathway. The infections are difficult to treat and therapeutic options are limited. We explored the feasibility of antisense-mediated exon skipping as therapy for MSMD with cells from a complete IL-12R
1-/- patient. Expression constructs were first studied to determine whether IL12RB1 lacking exon two encodes a functional protein. The IL-12R1 expression construct lacking exon two was expressed on T-cells. Upon IL-12 or IL-23 stimulation this construct phosphorylated similar amounts of STAT1, STAT3 and STAT4 and induced similar amounts of IFN- compared to a normal IL-12R1 construct. Antisense oligonucleotides (AONs) directed at exon two resulted in transcripts lacking exon two in both controls' and patients' T-cells. In IL-12R1-/- cells skipping of exon two lead to expression of IL-12R1 on the cell surface and responsiveness to IL-12. In conclusion, we showed that IL12RB1 lacking exon two encodes a functional IL-12R1. We demonstrated that T-cells can be highly efficiently transduced with AONs and are amenable to antisense-mediated exon skipping. Furthermore, we showed that exon skipping (partly) corrects the IL-12R1 deficiency in patients' cells.
